Advertisement

Immunophenotypic Analysis of CAR-T Cells

  • Júlia Teixeira Cottas de AzevedoEmail author
  • Amanda Mizukami
  • Pablo Diego Moço
  • Kelen Cristina Ribeiro Malmegrim
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 2086)

Abstract

CAR-T cell immunotherapy is a promising therapeutic modality for cancer patients. The success of CAR-T cell therapy has been associated with the phenotype, activation and functional profiling of infused CAR-T cells. Therefore, immunophenotypic characterization of CAR-T cells during bioprocess is crucial for cell quality control and ultimately for improved antitumor efficacy. In this chapter, we propose a flow cytometry panel to characterize the immunophenotype of the CAR-T subsets.

Key words

CAR-T cells Immunophenotypic analysis Flow cytometry T cells subsets Naive T cell Central memory T cell Effector memory T cell Effector T cell Memory stem T cells 

References

  1. 1.
    Saudemont A, Jespers L, Clay T (2018) Current status of gene engineering cell therapeutics. Front Immunol 9:153–167CrossRefGoogle Scholar
  2. 2.
    Porter DL, Levine BL, Kalos M et al (2011) Chimeric antigen receptor-modified T cells in chronic lymphoid leukemia. N Engl J Med 365(8):725–733CrossRefGoogle Scholar
  3. 3.
    Davila ML, Riviere I, Wang X et al (2014) Efficacy and toxicity management of 19-28z CAR-T cell therapy in B cell acute lymphoblastic leukemia. Sci Transl Med 6(224):224ra25CrossRefGoogle Scholar
  4. 4.
    Brentjens RJ, Davila ML, Riviere I et al (2013) CD19-targeted T cells rapidly induce molecular remissions in adults with chemotherapy-refractory acute lymphoblastic leukemia. Sci Transl Med 5(177):177ra38CrossRefGoogle Scholar
  5. 5.
    Liadi I, Singh H, Romain G et al (2015) Individual motile CD4(+) T cells can participate in efficient multikilling through conjugation to multiple tumor cells. Cancer Immunol Res 3(5):473–482CrossRefGoogle Scholar
  6. 6.
    Sommermeyer D, Hudecek M, Kosasih PL et al (2016) Chimeric antigen receptor-modified T cells derived from defined CD8+ and CD4+ subsets confer superior antitumor reactivity in vivo. Leukemia 30(2):492–500CrossRefGoogle Scholar
  7. 7.
    Louis CU, Savoldo B, Dotti G et al (2011) Antitumor activity and long-term fate of chimeric antigen receptor-positive T cells in patients with neuroblastoma. Blood 118(23):6050–6056CrossRefGoogle Scholar
  8. 8.
    Wang X, Popplewell LL, Wagner JR et al (2016) Phase 1 studies of central memory-derived CD19 CAR T-cell therapy following autologous HSCT in patients with B-cell NHL. Blood 127(24):2980–2990CrossRefGoogle Scholar
  9. 9.
    Gattinoni L, Lugli E, Ji Y et al (2011) A human memory T cell subset with stem cell-like properties. Nat Med 17(10):1290–1297CrossRefGoogle Scholar
  10. 10.
    Lugli E, Gattinoni L, Roberto A et al (2013) Identification, isolation and in vitro expansion of human and nonhuman primate T stem cell memory cells. Nat Protoc 8(1):33–42CrossRefGoogle Scholar
  11. 11.
    Gattinoni L, Klebanoff CA, Restifo NP (2012) Paths to stemness: building the ultimate antitumour T cell. Nat Rev Cancer 12(10):671–684CrossRefGoogle Scholar
  12. 12.
    Biasco L, Scala S, Basso Ricci L et al (2015) In vivo tracking of T cells in humans unveils decade-long survival and activity of genetically modified T memory stem cells. Sci Transl Med 7(273):273ra13CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2020

Authors and Affiliations

  • Júlia Teixeira Cottas de Azevedo
    • 1
    • 2
    • 3
    Email author
  • Amanda Mizukami
    • 1
  • Pablo Diego Moço
    • 1
  • Kelen Cristina Ribeiro Malmegrim
    • 4
  1. 1.Center for Cell-based Therapy, Regional Blood Center of Ribeirão Preto, Ribeirão Preto Medical SchoolUniversity of São PauloRibeirão PretoBrazil
  2. 2.Department of Biochemistry and Immunology, Ribeirão Preto Medical SchoolUniversity of São PauloRibeirão PretoBrazil
  3. 3.Department of Diabetes Immunology, Diabetes and Metabolism Research Institute at the Beckman Research Institute of City of Hope, DuarteCAUSA
  4. 4.School of Pharmaceutical Sciences of Ribeirão PretoUniversity of São PauloSão PauloBrazil

Personalised recommendations